激光扫描共聚焦显微镜观察瘦素在小鼠日本血吸虫病肝纤维化中的作用  

Laser scanning confocal microscope observes roles of leptin in liver fibrosis of mice with schistosomiasis japonica

作　　者：易芬兰[1] 牛丽文[2] 陶强[3] 冯亮[2] 邓庆强[2] 

机构地区：[1]江西省儿童医院科教科,南昌330006 [2]江西省儿童医院普外科,南昌330006 [3]江西省儿童医院新生儿外科,南昌330006

出　　处：《中华实验外科杂志》2016年第4期972-974,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察小鼠感染日本血吸虫后不同时期肝脏瘦素长受体（OB—Rb）蛋白动态表达，探讨瘦素在日本血吸虫病肝纤维化发生发展中的作用及意义。方法日本血吸虫尾蚴皮肤敷贴法感染小鼠构建日本血吸虫病肝纤维化模型，于感染后不同时间取肝脏标本。苏木素-伊红（HE）和VanGieson（VG）染色对肝组织进行病理学检查，免疫组织化学链霉菌抗生物素蛋白-过氧化物酶（SP）法动态观察OB—Rb及肝星状细胞（HSC）标志物α-平滑肌动蛋白（α-SMA）表达；激光扫描共聚焦显微镜（LSCM）联合免疫荧光双重标记技术进一步观察肝纤维化组织和虫卵OB-Rb和α-SMA蛋白表达。结果α-SMA和OB—Rb阳性表达出现的时间、部位和进程类似，随病程发展呈进行性加重，两者共表达于肝纤维化胶原纤维沉积处。结论HSC是日本血吸虫病肝纤维化发生发展过程中的关键细胞，瘦素在该过程中发挥重的促肝纤维化作用，可望成为新的治疗靶点。联合应用LSCM和免疫荧光双重标记技术能对蛋白进行精确的定位研究。Objective To observe the dynamic changes in the long form of leptin receptor （ OB - Rb） protein expression in mice liver by Schistosoma japonicum infection at different period, and to investigate the roles of leptin in liver fibrosis of mice with Schistosomiasis japonica. Methods Mice infec- ted with Schistosoma japonicum cercariae percutaneously were served as animal models of liver fibrosis, and live biopsies were done at different time point after infection. Liver lesions were evaluated using hematoxy- lin and eosin （HE） and Van Gieson （VG） staining. Immunohistochemistry for OB -Rb and α-smooth muscle actin （ α- SMA） as an activated hepatic stellate cell （HSC） marker was performed to observe their dynamic changes by streptavid - in - peroxidase （SP） technique. Laser Scanning Confocal Microscope （LSCM） combined with double label immunofluorescence technique observes the expression of OB - Rb and α- SMA in liver fibrosis and Schistosoma japonicum eggs. Results The expression time, location, and course of OB - Rb positive cells were similar to those of α- SMA positive cells. They were all located in collagen fiber deposition in liver fibrosis, increasing or aggravating progressively with course of diseases. Conclusion HSC plays an essential role in the formation and development of schistosomal live fibrosis, while leptin promote hepatic fibrogenesis in the process, which is expected to be a novel therapeutic target. Protein can be precisely targeted by LSCM combined with double label immunofluorescence technique.

关 键 词：日本血吸虫病 肝纤维化 肝星状细胞 瘦素 激光扫描共聚焦显微镜 

分 类 号：R532.210.6[医药卫生—内科学]